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Somatic Cell and Molecular Genetics

, Volume 15, Issue 3, pp 255–263 | Cite as

Regulation of tyrosinase mRNA levels in mouse melanoma cell clones by melanocyte-stimulating hormone and cyclic AMP

  • George E. Hoganson
  • Florence Ledwitz-Rigby
  • Richard L. Davidson
  • Bryan B. Fuller
Article

Abstract

Mouse melanoma cells in culture respond to melanocyte-stimulating hormone (MSH) by demonstrating increased activity of tyrosinase, the rate-limiting enzyme for melanin synthesis. Because this stimulation is strictly dependent upon continued transcription and translation, we have carried out studies to determine if MSH increases the level of tyrosinase mRNA. The abundance of tyrosinase message levels in melanoma cells treated with either MSH or dibutyryl cAMP was determined by Northern blot analysis utilizing a 946 base pair mouse tyrosinase cDNA probe. The tyrosinase cDNA was isolated from a λgt11 expression library generated from mRNA isolated from theopylline-induced Cloudman melanoma cells. The abundance of tyrosinase mRNA was determined in an amelanotic cell clone (AM-7AS) and a melanotic cell clone (MEL-11AS). The melanotic cell line had five times as much tyrosinase activity and almost 10 times more tyrosinase mRNA than the amelanotic line. Tyrosinase activity and mRNA increased in both cell lines after MSH addition. The amelanotic line treated with MSH for three days showed a fivefold increase in tyrosinase activity and a twofold increase in tyrosinase mRNA. The melanotic cell line treated with MSH for three days showed a 3.7-fold increase in enzyme activity and an eightfold increase in the abundance of tyrosinase mRNA. Dibutyryl cAMP also stimulated tyrosinase activity and the accumulation of tyrosinase mRNA. The data suggest that MSH, acting through cAMP, promotes an accumulation of tyrosinase mRNA.

Keywords

Melanoma Cell Cell Clone Northern Blot Analysis Tyrosinase Activity Melanin Synthesis 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Literature cited

  1. 1.
    Wong, G., and Pawelek, J. (1973).Nature (London) New Biol. 241:213–215.Google Scholar
  2. 2.
    Fuller, B.B., and Lebowitz, J. (1980).J. Cell Physiol. 103:279–287.PubMedGoogle Scholar
  3. 3.
    Pawelek, J., Wong, G., Sansone, M., and Morowitz, J. (1973).Yale J. Biol. Med. 46:430–443.PubMedGoogle Scholar
  4. 4.
    Fuller, B.B., and Viskochil, D.H. (1979).Life Sci. 24:2405–2416.PubMedGoogle Scholar
  5. 5.
    Fuller, B.B., Lunsford, J.B., and Iman, D.S. (1987).J. Biol. Chem. 262:4024–4033.PubMedGoogle Scholar
  6. 6.
    Jimenez, M., Kameyama, K., Maloy, W.L., Tomita, Y., and Hearing, V.J. (1988).Proc. Natl. Acad. Sci. U.S.A. 85:3830–3834.PubMedGoogle Scholar
  7. 7.
    Yamamoto, H., Takeuchi, S., Kudo, T., Makino, K., Nakata, A., Shinoda, T., and Takeuchi, T. (1987).Jpn. J. Genet. 62:271–274.Google Scholar
  8. 8.
    Kwon, B.S., Wakulchik, M., Haq, A.K., Halaban, R., and Kestler, D. (1988).Biochem. Biophys. Res. Commun. 153:1301–1309.PubMedGoogle Scholar
  9. 9.
    Müller, G., Ruppert, S., Schmid, E., and Schütz, G. (1988).EMBO J. 7:2723–2730.PubMedGoogle Scholar
  10. 10.
    Kwon, B.S., Haq, A.K., Pomerantz, S.H., and Halaban, R. (1987).Proc. Natl. Acad. Sci. U.S.A. 84:7473–7477.PubMedGoogle Scholar
  11. 11.
    Ruppert, S., Müller, G., Kwon, B., and Schütz, G. (1988).EMBO J. 7:2715–2722.PubMedGoogle Scholar
  12. 12.
    Fuller, B.B., Iman, D.S., and Lunsford, J.B. (1988).J. Cell Physiol. 134:149–154.PubMedGoogle Scholar
  13. 13.
    Chu, M.Y., and Fischer, G.A. (1968).Biochem. Pharmacol. 17:753–767.PubMedGoogle Scholar
  14. 14.
    Pomerantz, S.H. (1966).J. Biol. Chem. 241:161–168.PubMedGoogle Scholar
  15. 15.
    Bradford, M. (1976).Anal Biochem. 72:248–252.PubMedGoogle Scholar
  16. 16.
    Ross, J. (1976).J. Mol. Biol. 106:403–420.PubMedGoogle Scholar
  17. 17.
    Gubler, U., and Hoffman, B. (1983).Gene 25:263–269.PubMedGoogle Scholar
  18. 18.
    Maniatis, T., Fritsch, E.F., and Sambrook, J. (1982).Molecular Cloning: A Laboratory Guide, (Cold Spring Harbor Laboratory, New York).Google Scholar
  19. 19.
    Norrander, J., Kempe, T., and Messing, J. (1983).Gene 26:101–106.PubMedGoogle Scholar
  20. 20.
    Sanger, F., Nicklen, S., and Coulson, A.R. (1977).Proc Natl. Acad. Sci. U.S.A. 74:5463–5467.PubMedGoogle Scholar
  21. 21.
    Chomczynski, P., and Sacchi, N. (1987).Anal. Biochem. 162:156–159.PubMedGoogle Scholar
  22. 22.
    Fort, P., Marty, L., Piechaczyk, M., Sabrouty, S.E., Dani, C., Jeanteur, P., and Blanchard, J.M. (1985).Nucleic Acids Res. 13:1431–1442.PubMedGoogle Scholar
  23. 23.
    Cleveland, D.W., Lopata, M.A., MacDonald, R.J., Cowan, N.J., Rutter, W.J., and Kirschner, M.W. (1980).Cell 20:95–105.PubMedGoogle Scholar
  24. 24.
    Dexter, T.J., and Bennett, D.C. (1987).Exp. Cell Res. 168:255–264.PubMedGoogle Scholar
  25. 25.
    Lee, T.H., Lee, M.S., and Lu, M. (1972).Endocrinology 91:1180–1188.PubMedGoogle Scholar
  26. 26.
    Vijayan, E., Husain, I., Ramaiah, A., and Madan, N.C. (1982).Arch. Biochem. Biophys. 217:738–747.PubMedGoogle Scholar
  27. 27.
    Wong, G., and Pawelek, J. (1975).Nature 255:644–646.PubMedGoogle Scholar
  28. 28.
    Korner, A., and Pawelek, J. (1977).Nature 267:444–447.PubMedGoogle Scholar
  29. 29.
    Halaban, R., Pomerantz, S.H., Marshall, S., and Lerner, A.B. (1984).Arch. Biochem. Biophys. 230:383–387.PubMedGoogle Scholar

Copyright information

© Plenum Publishing Corporation 1989

Authors and Affiliations

  • George E. Hoganson
    • 1
  • Florence Ledwitz-Rigby
    • 2
  • Richard L. Davidson
    • 3
  • Bryan B. Fuller
    • 4
  1. 1.Department of PediatricsUniversity of Illinois, College of MedicineChicago
  2. 2.Department of Biological SciencesNorthern Illinois UniversityDeKalb
  3. 3.Department of GeneticsUniversity of Illinois, College of MedicineChicago
  4. 4.Department of Biochemistry and Molecular Biology, Department of DermatologyUniversity of Oklahoma Health Sciences CenterOklahoma City

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